Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 91, Issue 3, Pages 731-740Publisher
SPRINGER
DOI: 10.1007/s00253-011-3297-0
Keywords
Trimer motif; Mammalian cell; Signal peptide; Tissue plasminogen activator (tPA); Protein expression
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Funding
- National Natural Science Foundation of China [30771910]
- Ministry of Health of China [2008ZX10001-012]
- Natural Science Foundation of Heilongjiang Province [ZJY0601-01]
- Heilongjiang Innovation Program in Graduate Education [YJSCX2009-222HLJ]
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Ideal immunogenicity in antigens is a prerequisite to eliciting a sufficiently strong immune and memory response via either DNA or protein vaccines. To improve immunogenicity, efforts have focused on high-level expression of target proteins and on maintaining their natural conformations. In the present work, two trimer motifs (MTQ and MTI) were designed and introduced into a plasmid vector with the tissue plasminogen activator signal peptide (tPA-SP). Next, we examined the efficacy and the efficiency of the two motifs as well as the introduction of tPA-SP and its mutant forms, 22P/A and 22P/G, in facilitating the secretory expression of trimeric proteins in mammalian cells. We found that both trimer motifs could produce the target protein in a trimeric form at a high level. Introduction of tPA-SP 22P/A markedly increased the secretory expression level. The combination of the trimer motif, MTQ, and the signal peptide, 22P/A, may serve as a universal mammalian vector for producing trimeric proteins in vaccine development.
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